Method for treating wounds, burns and dermatological damage by administering selegiline or desmethylselegiline

ABSTRACT

The present invention is directed to methods that can be used in the treatment of wounds, burns, and photodamaged skin. Methods can be used for both humans and animals and involve the administration of compositions containing selegiline and/or desmethylselegiline.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims priority to international patentapplication number PCT/US99/04588, filed Mar. 3, 1999, which claimspriority to provisional patent application serial No. 60/078,043, filedMar. 16, 1998.

FIELD OF THE INVENTION

The present invention relates to methods for treating wounds, burns, ordermatological damage by administering topical compositions containingselegiline and/or desmethylselegiline.

BACKGROUND OF THE INVENTION

A. Free Radicals

Free radicals are molecules with one or more unpaired electrons in theirouter orbitals. The presence of these electrons together with thetendency of molecules to seek the lowest stable energy state causes freeradicals to be highly reactive and generally short lived. Among the freeradicals commonly found in vivo are oxygen, the superoxide anion and thehydroxyl radical. These are typically referred to as “oxidants” and areoften the result of cascades in which electrons are passed from moleculeto molecule.

B. Injuries and Free Radical Damage

Injuries such as wounds and burns generate free radicals that have bothlocal and systemic effects. Locally, free radicals have been implicatedin both tissue ischemia (Granger, et al., Gastroenterology 81:22-29(1981); Parks, et al., Gastroenterology 82:9-15 (1982)) and reperfusioninjuries (Schiller, et al., Critical Care Med. 21:S92-S100 (1993)).Systemically, burns often cause dysfunction of the heart, lungs andliver. Researchers have found that burn healing is improved when lipidperoxidation (typically caused by the action of free radicals) isreduced (LaLonde, et al., J. Burns Care & Rehabilitation 17:379-383(1996)).

C. Photodamage

Exposure of skin to electromagnetic radiation in the ultraviolet andvisible portions of the spectrum and ionizing radiation may result indamage to both the proteins and the DNA in skin cells. Such“photodamage” has been correlated with the induction of non-melanomaskin cancer, immune function suppression and photoaging.

Exposure of skin to ultraviolet and ionizing radiation and theconcomitant pathobiologies have been linked to the generation ofoxidants as well as to a reduction in anti-oxidant levels and activity(Stewart, et al., J. Inv. Dermatol. 106:1086-1089 (1996); Darr, et al.,Brit. J. Dermatol. 127:247-253 (1992)). Specifically, research has shownthat there is a reduction in epidermal superoxide dysmutase activity andin the levels of vitamin C and vitamin E after exposure to UVBradiation. Elimination of oxidants (e.g., by application of exogenousanti-oxidants) or prevention of oxidant production (e.g., by reductionof exposure to ionizing radiation) can alleviate or preventdermatological damage. The adverse effects of ionizing radiation includeedema, vasodilation, lymphocytic and neutrophilic infiltration in thedermis, dyskeratotic keratinocytes and spongiosis of the epidermis.

D. Use of Anti-Oxidants to Detoxify Free Radicals

A number of different strategies have been used in attempting to preventor reduce free radical damage. Endogenous anti-oxidants, e.g.,superoxide dysmutase, catalase or glutathione peroxidase, may be used toprotect cell membranes and agents such as ascorbic acid and glutathionemay be used to protect cytosols. Other anti-oxidants, such asalpha-tocopherol and tretinoin, have also been used to ameliorate theeffect of free radicals.

Administration of superoxide dysmutase, post-ischemia prevents theincreased capillary permeability which accompanies reperfusion injuries(Granger, et al., Gastroenterology 81:22-29 (1981)) and the ablation offree radical generation prior to, and at the time of, reperfusion mayprevent or lessen the severity of multiple system organ failure syndrome(Schiller, et al., Critical Care Med. 21:S92-S100 (1993)). Stewart, etal. have shown that UVB-induced DNA damage in human keratinocytes isattenuated by supplementing culture medium surrounding the cells withanti-oxidants such as vitamin C, selenite, or a water-soluble vitamin Eanalog (J. Inv. Dermatol. 106:1086-1089 (1996)).

E. Selegiline and Desmethylselegiline

Monoamine oxidase A (MAO-A) and monoamine oxidase B (MAO-B) are enzymesfound in both in the central nervous system and in peripheral tissues.MAO-A and MAO-B catalyze the oxidative deamination of primary amines,including neuroactive and vasoactive amines, resulting in the formationof toxic free radical species and free radical-generating cascades.Selegiline is a potent and selective inhibitor of monoamine oxidase Band has been reported to have an action in protecting or rescuingneurons of the central nervous system (Knoll, Mount Sinai J. Med.55:67-74 (1988)). Although the exact mechanism by which selegilinecauses its effects is not known, there is evidence suggesting that itmay provide neuroprotection or neuronal rescue by reducing oxidativedamage caused by monoamine oxidase and/or other oxidants (Jenner, etal., Neurology 47:S162-S170 (1996)). In this regard, selegiline has beenshown to increase the activity of the endogenous anti-oxidantssuperoxide dysmutase, catalase and glutathione peroxidase (Id.).

Desmethylselegiline, one of the metabolites of selegiline, exhibitsreduced MAO-B inhibitory activity in comparison to selegiline and itsactivity with respect to the inhibition of MAO-A is decreased to an evengreater extent. Thus, it is expected that desmethylselegiline shouldproduce selegiline-like neuroprotective effects with a decreased risk ofside effects associated with MAO-A inhibition.

Although selegiline has been used to treat Parkinson's disease, its useas a treatment for injuries, such as burns and wounds, and foralleviating dermatological damage, such as photodamage, has not beenknown heretofore. The present invention is directed to methods whichrely upon the administration of selegiline or desmethylselegiline tospeed the healing and reduce the complications associated with theseconditions.

SUMMARY OF THE INVENTION

The present invention is based upon the discovery that compositionscomprising selegiline and/or desmethylselegiline can be used in thetreatment of wounds, burns and photodamaged skin. In the case of burnsand wounds, compositions should be administered for a durationsufficient to promote epithelization. In the case of photodamaged skin,the composition should be administered for a duration sufficient topromote healing, as evidenced by a reduction in one or more of thesymptoms associated with photodamaged skin. These symptoms includeedema, vasodilation, lymphocytic and neutrophilic infiltration in thedermis, dyskeratotic keratinocytes and spongiosis of the epidermis.

Although the invention encompasses administration by any route, deliveryby means of a topical composition containing between 1×10−11 moles/literand 1×10×3 moles/liter of selegiline and/or desmethylselegiline ispreferred. Topical compositions may be delivered by means of a spray,patch, salve, cream, lotion or gel. As used herein, the term“desmethylselegiline” refers to either the R(−) enantiomeric form of thedrug, the S(+) enantiomeric form of the drug, or a racemic mixture ofthe two. In carrying out the present methods, the R(−) enantiomer may beused in the substantial absence of the S(+) enantiomer or vice versa. Anenantiomer is substantially absent if it constitutes less than 10% ofthe combined desmethylselegiline enantiomers. Compositions may containwater, suspending agents, thickeners, humectants, preservatives,emollients, emulsifiers and film formers. They may be applied eitherdirectly to the skin of a patient or they may be applied as part of apatch.

Although not preferred, non-topical routes of administration arecompatible with the present invention and may be used. The dosage ofselegiline or desmethylselegiline when used non-orally should be atleast 0.015 mg per kg body weight, calculated on the basis of the freesecondary amine, with progressively higher doses being employeddepending upon the route of administration and the subsequent responseto therapy. Typically, the daily non-oral dose will be about 0.10 mg/kgand may extend to about 1.0 mg/kg (all such doses again being calculatedon the basis of the free secondary amine).

DETAILED DESCRIPTION OF THE INVENTION

In the following description, reference will be made to variousmethodologies well known to those skilled in the art of medicine andpharmacology. Such methodologies are described in standard referenceworks setting forth the general principles of these disciplines. Unlessotherwise indicated, the descriptions apply to selegiline, and allenantiomeric forms of desmethylselegiline.

Dosage

The optimal daily dose of selegiline and/or desmethylselegiline usefulfor the purposes of the present invention may be determined by methodsknown in the art based upon clinical conditions such as the severity ofthe injury, the condition of the subject to whom treatment is beinggiven, the desired therapeutic response and the concomitant therapiesbeing administered to the patient or animal. Ordinarily, however, it isexpected that the attending physician or veterinarian will apply atopical composition containing a concentration of selegiline and/ordesmethylselegiline between 1×10−11 moles/liter and 1×10−3 moles/liter,preferably between 1×10−9 moles/liter and 1×10−3 moles/liter. Sufficientcomposition should be administered to completely cover the damaged areaon the individual's skin.

If the physician chooses non-oral routes of administration, at least0.015 mg/kg of selegiline and/or desmethylselegiline should beadministered daily with the more typical dosage being about 0.10 mg/kg.The daily dosage may be increased up to about 1.0 mg/kg. In all cases,doses are calculated on the basis of the free secondary amine form ofthe agent being administered. These guidelines further require that theactual dosage be carefully titrated by the attending physician orveterinarian depending upon the age, weight, clinical condition andobserved response of the individual being treated.

Topical compositions can be applied several times during the day towounded, burned or photodamaged skin. Similarly, daily dosages ofnon-oral preparations may be administered in a single or multiple dosageregiment. In addition, dosage forms permitting the continuous release ofactive agent, e.g., a transdermal patch, may be used for deliveringdrug.

Dosage Forms Route of Administration

As noted above, topical administration and topical dosage forms aregenerally preferred for the present methods. However, any of thenumerous dosage forms described in the literature for the administrationfor selegiline may be used and may include desmethylselegiline asdesired. For example, U.S. Pat. No. 4,812,481 discloses the use ofselegiline, in combination with amantadine, in oral, pectoral, internal,pulmonary, rectal, nasal, vaginal, lingual, intravenous, intraarterial,intracardial, intramuscular, intraperitoneal, intracutaneous, andsubcutaneous formulations. Dosage forms for selegiline having an outerwall with one or more pores in the wall impermeable to selegiline butpermeable to external fluids have been described. This dosage form mayhave applicability for oral, sublingual, or buccal administration.Similarly, a variety of selegiline compositions, including tablets,pills, capsules, powders, aerosols, suppositories, skin patches,parenterals, and oral liquids, including oil suspensions, solutions andemulsions have been described. Further disclosed areselegiline-containing sustained release (long acting) formulations anddevices.

Topical dosage forms may be prepared according to conventionaltechniques with creams being generally preferred. The topical cream maybe a cosmetically elegant oil in water, cream/lotion/emulsion,containing the desired specified concentration of selegiline and/ordesmethylselegiline. Such moisturizing cream formulations may contain avehicle, a buffer system to maintain the vehicle at an appropriate pH,and an acceptable antimicrobial preservative system. The cream mayfurther contain thickeners, humectants, emollients, emulsifiers and filmformers. Methods for preparing appropriate formulations are well knownin the art (see e.g., Remington's Pharmaceutical Sciences, 16th ed., A.Oslo. ed., Easton Pa. (1980)).

Transdermal dosage forms can be prepared utilizing a variety oftechniques that have been described in the art. Examples may be found inU.S. Pat. Nos. 4,861,800; 4,868,218; 5,128,145; 5,190,763; and5,242,950; and in foreign patent documents EP-A 404807; EP-A 509761; andEP-A 593807. A monolithic patch structure can be utilized in which drugis directly incorporated into the adhesive and this mixture is cast ontoa backing sheet. EP-A 593807 describes a composition in which selegilineis administered as an acid addition salt by incorporating it into amulti layer patch which promotes a conversion of the salt into the freebase form of selegiline. One can also employ a device using a lyotropicliquid crystalline composition in which, for example, 5-15% ofselegiline is combined with a mixture of liquid and solid polyethyleneglycols, a polymer and a non-ionic surfactant, optionally with theaddition of propylene glycol and an emulsifying agent. Further detailson the preparation of such transdermal formulations are found in thepatent literature.

Buccal and sublingual dosage forms of selegiline and/ordesmethylselegiline may be prepared utilizing techniques described in,for example, U.S. Pat. Nos. 5,192,550; 5,221,536; 5,266,332; 5,057,321;5,446,070; or 5,354,885.

Chemical Form of Selegiline or Desmethylselegiline

The present invention is not limited to any particular form ofselegiline and/or desmethylselegiline and drugs may be used either asfree bases or as pharmaceutically acceptable acid addition salts. In thelatter case, the hydrochloride salt is generally preferred. However,other salts derived from organic and inorganic acids may also be used.

Manner of Treatment

The methods disclosed herein may be used for both human and nonhumansubjects. With regard to the latter, the methods are particularly, butnot exclusively, directed to domesticated mammals such as canine andfeline species.

In the case of wounds and burns, treatment by administration ofselegiline and/or desmethylselegiline should be continued untilepithelialization is complete. For dermatological damage, treatmentshould be continued until the related symptoms, such as edema,vasodilation, lymphocytic and neutrophilic infiltration or spongiosis ofthe epidermis, subside. The drugs may be either administered at regularintervals (e.g., twice a day) or in an essentially continuous manner(e.g. via a transdermal patch).

EXAMPLES Example 1

Protective Effects of Selegiline and Desmethylselegiline AgainstPhotodamage

The ability of selegiline and/or desmethylselegiline to preventphotodamage can be correlated to the reduction of cell apoptosisfollowing exposure to UVB radiation. Previous research has shown thatprimary human keratinocytes grown in serum-free medium are susceptibleto UVB-induced apoptosis when exposed to 600-800 J/m2. In addition,deprivation of growth factors, specifically insulin, increaseskeratinocyte sensitivity to UVB radiation with apoptosis occurring atUVB levels of 200 J/m2.

Two sets of primary human keratinocytes are grown in complete, andgrowth factor deprived (GFD) media for a total of four sets of cultures.The keratinocytes are then placed in GFD or complete media containing 0,1×10−9, 1×10−8, 1×10−7, 1×10−5, 1×10−6, 1×10−4, and 1×10−3 M selegilineor desmethylselegiline. Twenty-four hours after the addition of thedrugs, the keratinocytes grown in GFD medium are irradiated with zero or200 J/m2 UVB and the keratinocytes grown in complete media areirradiated with zero or 800 J/m2 UVB. Using morphological examination,DAPI staining, Annexin V-FITC FACS analysis and PARP cleavage analysis,the keratinocytes are tested for apoptosis 15 hours after irradiation.

First, the keratinocytes are examined for morphological signs ofapoptosis. Following morphological examination, the cells are harvestedby trypsinization and pelleted by low speed centrifugation. Each pelletis divided into three parts for the remaining evaluation.

DAPI staining is used to examine the cells for nuclear condensationcharacteristic of cell apoptosis. An aliquot of each cell pellet iswashed with PBS, fixed in Histochoice, and resuspended in DAPI stainingsolution for one hour. The cells are subsequently washed in PBS andattached to microscope slides by cytospins. The cells are thenvisualized by epifluorescent microscopy and the percentage of apoptoticcells determined.

Annexin V-FIT FACS analysis is used to determine extracellularphosphatidylserine exposure. In the early stages of cell apoptosis,portions of the plasma membrane translocate causing the normallyintracellular phosphatidylserine to move to the extracellular surface ofthe plasma membrane. Annexin V binds to the extracellularphosphatidylserine and the amount of such binding is measured by flowcytometry. An aliquot of each cell pellet is washed in PBS andresuspended in a binding buffer. The cells are then incubated in thedark for 15 minutes in the presence of Annexin V-FIT and propidiumiodide. Following incubation, the percentage of apoptotic cells isdetermined using FACS analysis. Apoptotic cells are Annexin V-FACSpositive and propidium iodide negative.

PARP cleavage analysis is used to measure the proteolytic cascade whichoccurs during apoptosis. One of the substrates for apoptosis-relatedproteases is poly(ADP-ribose) polymerase, or PARP. After cleavage, PARPreduces to a characteristic 85 kD fragment. An aliquot of each cellpellet is resuspended in RIPA buffer containing 7% urea and proteinlysates. The resulting cell proteins are then transferred to Immobilin Pmembranes by semi-dry electrophoresis and incubated in TSB buffer (150mM sodium chloride; 100 mM Tris-base, pH 7.5; 2% blocking reagent B:Boehringer Mannheim) for 2 hours at room temperature. Anti-PARPmonoclonal antibodies (clone C-2-10) are diluted in TSB and 2% blockingreagent B. Following one hour of incubation, the membrane is washedthree times with TSB buffer containing 0.1% Tween20. After theincubation is completed, biotinylated goat (Fab′-fragments) anti-mouseIg diluted in TSB is added to the membrane and the membrane is thenwashed three times with TSB buffer containing 0.1% Tween20. Astreptavidin-horseradish peroxidase conjugate is added to the membraneand the protein bands are visualized using Enhanced ChemiluminescentPlus (Amersham). The percent of PARP cleavage is determined usingdensitometry to compare the total PARP protein with the cleaved PARPfragment (85 kD).

Example 2

Curative Effects of Selegiline and Desmethylselegiline on Burns

The ability of selegiline and desmethylselegiline to assist in thehealing of burns was tested using a second-degree burn wound model. Insecond-degree burns, the entire surface of the epidermis is destroyed.An epidermal covering is regenerated from the remaining epithelial andepidermal cells adjacent to the burns. This phase of the healing processis called epithelization.

A. Materials and Methods

Experimental Animals

Swine were used for the experimental research animal because their skinhas many morphological similarities to human skin. Seven young femalespecific pathogen free (SPF:Ken-O-Kaw Farms, Windsor, Ill.) pigsweighing 25-30 kg were kept in house for two weeks prior to initiatingthe experiment. These animals were fed a basal diet ad libitum andhoused individually in facilities with controlled temperature (19-21°C.) and lights (12 h/12 h LD).

Wounding Technique

Experimental animals were clipped with standard animal clippers on theday of the experiment. The skin on the back and both sides of the animalwas prepared for wounding by washing with a non-antibiotic soap andsterile water. Each animal was anesthetized i.m. with ketamine-HCl (20mg/kg), xylazine (2 mg/kg) and atropine (0.05 mg/kg), followed by maskinhalation of an isoflurane and oxygen combination. Five specificallydesigned cylindrical brass rods weighing 358 g each were heated in aboiling water bath to 100° C. A rod was removed from the water bath andwiped dry before being applied to the skin surface to prevent waterdroplets from creating a steam burn on the skin. The brass rod was heldat a vertical position on the skin for six seconds with all pressurebeing supplied by gravity in order to make a burn 8.5 mm diameter×0.8 mmdeep. Immediately after burning, the roof of the burn blister wasremoved with a sterile spatula. The burn wounds were made approximately2 cm from each other.

Approximately 90 burn wounds were made on the anterior two-thirds of theanimal. The posterior third of the animal cannot be used because ofanatomical differences in burn wound healing (a more rapid healing ofburns has been observed at that position). Burn wounds were randomlyassigned to seven treatment groups and were applied in the pattern shownin Table 1.

TABLE 1 Treatment Groups Number of Animals Treatment Groups 1 agent A(dose X) agent A (dose Y) agent A (dose Z) 1 agent A (dose Y) agent A(dose Z) agent B (dose X) 1 agent A (dose Z) agent B (dose X) agent D(dose Y) 1 agent B (dose X) agent B (dose Y) agent B (dose Z) 1 agent B(dose Y) agent B (dose Z) air exposed, control 1 agent A (dose X) agentB (dose Z) air exposed, control 1 agent A (dose X) agent A (dose Y) airexposed, control agent A = selegiline agent B = desmethylselegiline HCLdose X = 10⁻⁴ M dose Y = 10⁻⁶ M dose Z = 10⁻⁸ M

A total of seven animals were used and a total of fifteen wounds pertreatment group were analyzed on each of days 7-12 after wounding.Treatments were coded to maintain blind study compliance. The burnwounds were treated with 25 ug of test article (enough to cover eachwound). Treatments were allowed to penetrate into the sites for at leasta 20 minute time period. Each treatment was applied within 20 minutes ofblister removal and treatment occurred once a day for the first fivedays.

Epidermal Migration Assessment

Beginning on day seven after wounding (day 0) and each day thereafterfor four to six days, five burn wounds and the surrounding normal skinfrom each treatment area were excised using an electrokeratome. Anyspecimens that were not excised intact were discarded. The excisedwounds and the surrounding normal skin were incubated in 0.5 M NaBr for24 hours at 37° C. After incubation, the specimens were separated intoepidermal and dermal sheets. The epidermis was examined macroscopicallyfor defects in the area of the burn wounds. Epithelization wasconsidered complete if no defect was present (healed). Any defect in theburn area indicated that healing was incomplete. The epidermal sheet wasplaced on cardboard for a permanent record.

B. Result

After the study was completed, the codes were revealed and data wastabulated. The number of wounds healed (completely epithelized) weredivided by the total number of wounds sampled per day and multiplied by100. Results are shown in Table 2.

TABLE 2 Epithelization Results (combined data)* TREAT- DAYS AFTERBURNING MENT 7 8 9 10 11 12 13 Saline {fraction (0/15)} {fraction(0/15)} {fraction (0/15)} {fraction (0/15)} {fraction (5/15)} {fraction(6/15)} {fraction (10/15 )} Control (0%) (0%) (0%) (0%) (33%) (40%)(67%) A-1 {fraction (0/15)} {fraction (0/15)} {fraction (0/15)}{fraction (3/15)} {fraction (5/15)} {fraction (6/15)} {fraction (6/15)}(0%) (0%) (0%) (20%)  (33%) (40%) (40%) A-2 {fraction (0/15)} {fraction(0/15)} {fraction (0/15)} {fraction (3/15)} {fraction (5/15)} {fraction(6/15)} {fraction (6/15)} (0%) (0%) (0%) (20%)  (33%) (40%) (40%) A-3{fraction (0/15)} {fraction (0/15)} {fraction (0/15)} {fraction (0/15)}{fraction (0/15)} {fraction (1/15)} {fraction (6/16)} (0%) (0%) (0%)(0%)  (0%)  (7%) (38%) B-1 {fraction (0/15)} {fraction (0/15)} {fraction(0/15)} {fraction (0/15)} {fraction (1/15)} {fraction (5/15)} {fraction(11/16 )} (0%) (0%) (0%) (0%)  (7%) (33%) (69%) B-2 {fraction (0/15)}{fraction (0/15)} {fraction (0/15)} {fraction (0/15)} {fraction (5/15)}{fraction (7/15)} {fraction (6/16)} (0%) (0%) (0%) (0%) (33%) (47%)(100%)  B-3 {fraction (0/15)} {fraction (0/15)} {fraction (0/15)}{fraction (0/15)} {fraction (5/15)} {fraction (10/15 )} {fraction(40/15 )} (0%) (0%) (0%) (0%) (33%) (67%) (67%) *Wounds are presented asthe number of wounds healed (completely epithelized) over the number ofwounds assessed. () Percent of wounds completely epithelized Code: A =selegiline HCL; B = desmethylselegiline HCL 1 = 10-4M 2 = 10-6M 3 =10-8M

All wounds absorbed the administered compounds within 10 to 15 minutesof application. During the first three days of treatment, the woundcrust in all treatment groups turned a white color during absorption ofapplied agent. This was more pronounced with treatments A and B. After15 minutes, the crust's color became normal again. Neither residue,erythema or infection were observed in any treatment groups. The resultsshown in Table 2 may be summarized as follows:

Day 7-9: None of the burn wounds were completely epithelized.

Day 10: Twenty percent (20%) of wounds which were treated withselegiline HCl at concentrations of 1×10−4 M and 1×10−6 M werecompletely epithelized. None (0%) of the wounds from any other treatmentgroups were completely epithelized.

Day 11: Thirty-three percent (33%) of wounds treated with eitherselegiline HCl (10−4 M and 10−6M), desmethylselegiline HCl (10−6 M and10−8M), or saline were completely epithelized. Seven percent (7%) of thedesmethylselegiline HCl(10−4M) treated wounds were completelyepithelized. None of the wounds treated with selegiline HCl(10−8 M) werecompletely epithelized.

Day 12: Thirty-three percent (33%) and forty percent (40%) of woundstreated with selegiline HCl(10−4 M and 10−6 M respectively), werecompletely epithelized. Only seven percent (7%) of wounds treated withthe 10−8M concentration of selegiline HCl were completely epithelized.Thirty-three percent (33%), forty-seven percent (47%) and sixty-sevenpercent (67%) of wounds treated with desmethylselegiline HCl (10−4M and10−6M and 10−8M respectively) were completely epithelized. Forty percent(40%) of wounds treated with saline, i.e., the control cells, werecompletely epithelized.

Day 13: Thirty-three (33%) and forty (40%) percent of wounds treatedwith selegiline HCl (10−4M and 10−6M respectively) were completelyepithelized. Thirty-eight percent of wounds treated with the 10−8 Mconcentration of selegiline HCl were completely epithelized. Sixty-nine(69%), one hundred (100%) and sixty-seven (67%) percent of woundstreated with desmethylselegiline HCl (10−4 M, 10−6 M, and 10−8 Mrespectively) were completely epithelized. Sixty-seven (67%) percent ofwounds treated with saline, i.e., controls, were completely epithelized.

C. Discussion

The data from these studies suggest that the wounds treated withselegiline HCl (10−4 M and 10−6 M) were able to initiate epithelizationat an earlier time point than all other treatment groups. However,wounds treated with desmethylselegiline HCl (all concentrations) had ahigher percentage of wounds completely epithelized on day thirteen.Wounds treated with desmethyl-selegiline HCl (10−6 M) were completelyepithelized sooner than all other treatment groups.

Example 3

Use of Selegiline and Desmethylselegiline to Improve Appearance ofPhotodamaged Skin

The test subjects, human females exhibiting moderate to severephotoaging on the dorsal portions of their forearms and hands, arerandomly assigned to receive either vehicle plus selegiline or vehiclealone. For a period of 16 weeks, the test subjects apply sufficientcream to cover the designated test area BID. Clinical assessments of theappearance of the test area are made prior to administration of thecream and at the second, eighth, sixteenth and twenty-fourth weeks.

Example 4

Use of Selegiline to Preserve the Positive Effects of Tretinoin onPhotoaged Skin

The test subjects, human females exhibiting moderate to severephotoaging on the face and dorsal portions of their forearms, arerandomly assigned to apply either the vehicle plus selegilinecombination or vehicle alone. For an initial period of sixteen weeks,all test subjects treat bilateral aspects of their face, forearms andhands with a tretinoin cream once per day. For a subsequent one weekperiod, all test subjects use no treatment on the test areas. For thefollowing sixteen week period, the test subjects apply cream, eithercontaining vehicle alone or vehicle plus selegiline to cover one half ofthe designated area BID. Clinical assessments of the test areas are madeprior to the administration of the cream and throughout the study.

All references cited herein are fully incorporated by reference. Havingnow fully described the invention, it will be understood by those ofskill in the art that the invention may be performed within a wide andequivalent range of conditions, parameters and the like, withoutaffecting the spirit or scope of the invention or any embodimentthereof.

What is claimed is:
 1. A method of treating a wound in a subjectcomprising administering a composition comprising selegiline and/ordesmethylselegiline to said subject in an amount effective to promoteepithelization of said wound.
 2. The method of claim 1, wherein saidcomposition is a topical composition comprising selegiline at aconcentration of between 1×10−11 moles/liter and 1×10−3 moles/liter. 3.The method of claim 1, wherein said composition is a topical compositioncomprising desmethylselegiline at a concentration of between 1×10−11moles/liter and 1×10−3 moles/liter.
 4. The method claim 1 wherein saidcomposition comprises desmethylselegiline in the form of its R(−)enantiomer and the S(+) enantiomer is substantially absent.
 5. Themethod of claim 1, wherein said composition comprisesdesmethylselegiline in the form of its S(+) enantiomer and the R(−)enantiomer is substantially absent.
 6. The method of either claim 2 or3, wherein said composition is in the form of a cream.
 7. The method ofeither claim 2 or 3, wherein said topical composition is in the form ofa gel.
 8. The method of claim 1, wherein said composition comprisesselegiline delivered by means of a patch.
 9. The method of claim 1,wherein said composition comprises desmethylselegiline delivered bymeans of a patch.
 10. The method of claim 1, wherein said compositionfurther comprises one or more compounds selected from the groupconsisting of water, suspending agents, thickeners, humectants,preservatives, emollients, emulsifiers, and film formers.
 11. A methodof treating a burn in a subject comprising administering a compositioncomprising selegiline and/or desmethylselegiline to said subject in anamount effective to promote epithelization of said burn.
 12. The methodof claim 11, wherein said composition is a topical compositioncomprising selegiline at a concentration of between 1×10−11 moles/literand 1×10−3 moles/liter.
 13. The method of claim 11, wherein saidcomposition is a topical composition comprising desmethylselegiline at aconcentration of between 1×10−11 moles/liter and 1×10−3 moles/liter. 14.The method of claim 11, wherein said composition comprisesdesmethylselegiline in the form of its R(−) enantiomer and the S(+)enantiomer is substantially absent.
 15. The method of claim 11, whereinsaid composition comprises desmethylselegiline in the form of its S(+)enantiomer and the R(−) enantiomer is substantially absent.
 16. Themethod of either claim 12 or 13, wherein said topical composition is inthe form of a cream.
 17. The method of either claim 12 or 13, whereinsaid topical composition is in the form of a gel.
 18. The method ofclaim 11, wherein said composition comprises selegiline delivered bymeans of a patch.
 19. The method of claim 11, wherein said compositioncomprises desmethylselegiline delivered by means of a patch.
 20. Themethod of claim 11, wherein said composition further comprises one ormore compounds selected from the group consisting of water, suspendingagents, thickeners, humectants, preservatives, emollients, emulsifiers,and film formers.